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Method for storing duodenal juice sample

A technique for duodenal juice and a preservation method is applied in the field of a kit for preparing the preservation sample, which can solve the problems of easy decomposition of protein markers, and achieve the effects of stable preservation and inhibition of decomposition

Active Publication Date: 2018-01-02
OLYMPUS CORP
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In particular, at the time of endoscopic examination, in order to observe the gastric mucosa, a strong protease for excreting the gastric mucosa is orally administered in advance, so the duodenal fluid collected during the endoscopic examination contains a very large amount of protease. Potent protease, there is a problem that protein markers are more easily broken down

Method used

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  • Method for storing duodenal juice sample
  • Method for storing duodenal juice sample
  • Method for storing duodenal juice sample

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0083] Duodenal fluid samples collected from four subjects (subjects a to d) were stored under various conditions, and the S100P concentration in the stored samples was measured. The measurement results were compared using the S100P concentration of a sample frozen immediately after collection and stored at -80°C as a reference.

[0084] Regarding the collection of the duodenal fluid sample from the test subject, a collection tool detachably provided with a container for storage containing 10 mg of powdered protease inhibitor (a mixture of AEBSF, PMSF, and TLCK) was passed through an endoscope. conduct. It should be noted that the length of time from the mixing time of the collected duodenal fluid sample and the protease inhibitor to the dilution with the ELISA buffer described later is about 1 to 3 minutes.

[0085] From the duodenal fluid samples mixed with protease inhibitors collected from each subject, take 20 μL to 6 containers respectively, 3 samples of which are direc...

Embodiment 2

[0091] For the duodenal fluid sample collected from subjects a to d in Example 1 mixed with 10 mg of powdered protease inhibitor (a mixture of AEBSF, PMSF, and TLCK), the effect of the component difference of the dilution solution on the S100P residue was investigated. rate (%).

[0092] Specifically, 20 μL to 12 containers were taken from the duodenal fluid samples mixed with protease inhibitors collected from each test subject, and 5 of them were filled with PBS containing 0.1% by mass of PBS (PBS+BSA0.1%) of BSA, PBS (PBS+BSA0.02%) containing 0.02 mass % BSA, PBS containing 0.1 mass % BSA and 0.05% tween (Tween) 20 (PBS +BSA 0.1%+tween 20 0.05%) or the ELISA buffer used in Example 1 were diluted to 100 times the volume, and stored at room temperature for 1 day. The remaining 6 samples were stored at -80°C for 1 day without dilution.

[0093] The S100P concentration (pg / mL) in the total amount of each sample after storage was measured in the same manner as in Example 1. H...

Embodiment 3

[0101] For the duodenal fluid sample collected from subjects a to d of Example 1 mixed with 10 mg of powdered protease inhibitor (a mixture of AEBSF, PMSF and TLCK), the effect of the dilution factor on the S100P residual rate (%) was investigated. Impact.

[0102]Specifically, 20 μL was taken from the duodenal fluid sample mixed with the protease inhibitor collected from each test subject to 4 containers, and the ELISA used in Example 1 was used for 3 samples The buffer solution was diluted to 5, 20, or 100 times volume, respectively, and then stored at room temperature for 1 day. The remaining one sample was stored at -80°C for 1 day without dilution.

[0103] Regarding the S100P concentration of each sample, the relative value (%) when the S100P concentration of the sample stored at -80°C was taken as 100% was determined as the S100P remaining rate (%). The results are shown in Table 4. As a result, for any test subject, the higher the dilution factor of the ELISA buffer...

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Abstract

The present invention addresses the problem of providing: a method for preparing a stock sample for stably storing a protein marker in a duodenal juice sample; and a kit for preparing the stock sample. The method comprises the steps of (a) diluting a duodenal juice sample collected from a subject with a diluting solution at a dilution factor of 10 folds by volume or more and (b) storing a dilutedduodenal juice sample, which is prepared in step (a), in a liquid form, wherein the diluting solution is an aqueous solution containing a protein at a concentration of 0.02% by mass or more.

Description

technical field [0001] The present invention relates to a method for preparing a preservation sample for stably preserving a protein marker in a duodenal fluid sample, and a kit for preparing the preservation sample. Background technique [0002] Duodenal juice is a mixture of pancreatic juice discharged from the pancreas, bile discharged from the bile duct, and mucus secreted from the duodenum. The collection of duodenal fluid is performed only by inserting an endoscope into the duodenum and suctioning at that location, which is less invasive and can be performed with simpler skills than pancreatic juice collection via the pancreatic duct . Therefore, pancreatic disease can be checked by detection of pancreatic disease markers in duodenal fluid. [0003] On the other hand, duodenal juice contains a lot of proteases from pancreatic juice and intestinal juice, and bacteria from bile and intestinal juice. Under the action of these, various biomolecules such as proteins conta...

Claims

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Application Information

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IPC IPC(8): G01N33/48C12M1/26G01N1/28G01N33/53
CPCC12M1/26G01N1/28G01N33/48G01N33/53
Inventor 佐贯博美中田真理
Owner OLYMPUS CORP
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